Journal: Journal of Clinical Medicine

Article Title: Th2 Cytokines Affect the Innate Immune Barrier without Impairing the Physical Barrier in a 3D Model of Normal Human Skin

doi: 10.3390/jcm12051941

Figure Lengend Snippet: Antibodies and protocols for indirect immunofluorescence analysis.

Article Snippet: Polyclonal rabbit anti-human ZO-1 (ThermoFisher Scientific) , Pronase E 10 min at 37°C , dilution 1:100 1 h at 37 °C.

Techniques: Immunofluorescence, Incubation

Journal: International Journal of Nanomedicine

Article Title: In vitro placental model optimization for nanoparticle transport studies

doi: 10.2147/IJN.S26601

Figure Lengend Snippet: BeWo monolayer (day 3 PS) grown on PC membranes with 3 μm pore size, showing ZO-1 tight junction protein staining. Notes: Blue, DAPI; Green, ZO-1 tight junction protein. Field of view 179 × 153 μm. Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; PS, postseeding.

Article Snippet: For transport studies using Fluoresbrite particles, DMEM-F12 without phenol red (Sigma-Aldrich, Irvine, Ayrshire, UK) supplemented with 10% (v/v) FBS (In Vitro, Fredensborg, Denmark), 1% (v/v) penicillin/streptomycin (Panum Institute, University of Copenhagen, Denmark) and 4 mM glutamine (In Vitro) was used, to avoid interference with optical absorbance; Hank’s balanced salt solution (HBSS), 4% (v/v) formaldehyde, dimethyl sulfoxide and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Sigma-Aldrich, Irvine, Ayrshire, UK); fluorescein sodium salt (Na-Flu), paraformaldehyde (PFA), Triton X-100 (Sigma-Aldrich, Dorset, UK), trypsin-ethylenediaminetetraacetic acid (EDTA; US Food and Drug Administration, Silver Spring, MD) and horse and goat sera (Sigma-Aldrich, Dorset, UK); primary polyclonal rabbit anti-human ZO-1 antibody (Zymed, South San Francisco, CA); Alexa Fluor ® 488 donkey antirabbit secondary antibody (Invitrogen, Paisley, UK); phalloidin (Sigma-Aldrich, Dorset, UK); VECTASHIELD mounting medium with 4′,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories Ltd, Peterborough, UK) for monolayer confirmation or VECTASHIELD mounting medium (Vector Laboratories Ltd, Burlingame, CA) for NP transport and uptake study; lysis solution (0.5% Triton X-100 in 0.2 M NaOH) (VWR, Herlev, DK).

Techniques: Staining

Journal: International Journal of Nanomedicine

Article Title: Uptake and transport of pullulan acetate nanoparticles in the BeWo b30 placental barrier cell model

doi: 10.2147/IJN.S161319

Figure Lengend Snippet: Confocal and TEM images of BeWo b30 cells. Notes: BeWo b30 cells were grown on PC membranes with 3 μm pore size. Cell nuclei were stained with DAPI (blue) and tight junctions with ZO-1 (green) ( A ). Monolayer formation and microvilli structure: tight junctions ( B ) and villus structure ( C ). Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; PC, polycarbonate; TEM, transmission electron microscopy; ZO-1, zonula occludens-1.

Article Snippet: Primary polyclonal rabbit anti-human zonula occludens-1 (ZO-1) antibody and Alexa Fluor-488 goat anti-rabbit secondary antibody were purchased from Thermo Fisher Scientific.

Techniques: Staining, Transmission Assay, Electron Microscopy